US4446122A
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Purified human prostate antigen
Assignee: RESEARCH Corp TECHNOLOGIES Inc A CORP OF
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Filing: 1980-12-23
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Granted: 1984-05-01
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Expires: 2000-12-23
Abstract
Abstract PCT No. PCT/US80/01708 Sec. 371 Date Aug. 28, 1981 Sec. 102(e) Date Aug. 28, 1981 PCT Filed Dec. 23, 1980 PCT Pub. No. WO81/01849 PCT Pub. Date Jul. 9, 1981.A prostate antigen distinct from prostatic acid phosphatase has been detected in normal, benign hypertrophic and malignant prostatic tissues, but not in other human tissues. The prostate antigen was purified to homogeneity from prostatic tissues by ammonium sulfate precipitation, DEAE-BioGel A anion exchange chromatography, molecular sievings on Sephadex G-100 and Sephadex G-75, and preparative polyacrylamide gel electrophoresis. The purified prostate antigen shows a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing. The molecular weight of purified antigen was estimated by Sephadex G-75 gel filtration to be 33,000 and by sodium dodecyl sulfate polyacrylamide gel electrophoresis to be 34,000 with no subunit. The prostate antigen had an isoelectric point of 6.9.
Inventors
- 👤 Tsann M. Chu
- 👤 Ming C. Wang
- 👤 Lawrence Papsidero
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-10-22 22:57
CPC Classifications
C; C07; C07K; C07K16/00; C07K16/18; C07K16/28; C07K16/30; C07K16/3038; A; A61; A61K; A61K39/00; A61K39/0005; A61K39/0011; A61K39/001193; A61K39/001194; G; G01; G01N; G01N33/00; G01N33/48; G01N33/50; G01N33/53; G01N33/531; G01N33/543; G01N33/558; G01N33/561; Y; Y10; Y10S; Y10S435/00; Y10S435/961; Y10S435/962
Claims Analysis
Total Claims: 19
Claim 1
Independent Claim
112 words
An in vivo composition of matter comprising an immunochemically effective concentration and amount of a purified human prostate specific antigen occurring in normal and cancerous prostatic tissue and purified to show a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing, said antigen having a molecular weight as determined by Sephadex G-75 gel of filtration of about 33,000 and by a molecular weight as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis of about 34,000 with no subunit and having an isoelectric point pI of 6.9, said antigen further being insoluble in perchloric acid and substantially free of normal serum protein components, water-insoluble cellular material and of prostatic acid phosphatases.
Claim 2
Dependent Claim
112 words
An in vivo composition of matter comprising an immunochemically effective concentration and amount of a purified human prostate specific antigen occurring in normal and cancerous prostatic tissue and purified to show a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing, said antigen having a molecular weight as determined by Sephadex G-75 gel of filtration of about 33,000 and by a molecular weight as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis of about 34,000 with no subunit and having an isoelectric point pI of 6.9, said antigen further being insoluble in perchloric acid and substantially free of normal serum protein components, water-insoluble cellular material and of prostatic acid phosphatases.
Claim 3
Dependent Claim
100 words
An in vitro composition of matter comprising an immunochemically effective concentration and amount of antibodies against a human prostate specific antigen occurring in normal and cancerous prostatic tissue and purified to show a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing, said antigen having a molecular weight by Sephadex G-75 gel filtration of about 33,000 and by sodium dodecyl sulfate polyacrylamide gel electrophoresis of about 34,000 with no subunit and having an isoelectric point pI of 6.9, said antigen further being insoluble in perchloric acid and said composition being free of antibodies against human prostatic acid phosphatases.
Claim 4
Dependent Claim
100 words
An in vitro composition of matter comprising an immunochemically effective concentration and amount of antibodies against a human prostate specific antigen occurring in normal and cancerous prostatic tissue and purified to show a single protein band on analytical polyacrylamide gel electrophoresis and isoelectric focusing, said antigen having a molecular weight by Sephadex G-75 gel filtration of about 33,000 and by sodium dodecyl sulfate polyacrylamide gel electrophoresis of about 34,000 with no subunit and having an isoelectric point pI of 6.9, said antigen further being insoluble in perchloric acid and said composition being free of antibodies against human prostatic acid phosphatases.
Claim 5
Dependent Claim
12 words
A composition according to claim 2, wherein said antibodies are monoclonal antibodies.
Claim 6
Dependent Claim
12 words
A composition according to claim 2, wherein said antibodies are immunoprecipitating antibodies.
Claim 7
Dependent Claim
16 words
A composition according to claim 2, wherein said antibodies are labeled for radioimmunoassay or enzyme-linked immunoassay.
Claim 8
Dependent Claim
16 words
A composition according to claim 2, wherein said antibodies are covalently bonded to a water-insoluble support.
Claim 9
Dependent Claim
75 words
A process for preparing immunoprecipitating antibodies to antigens associated with prostatic tissue and fluid, which comprises: (a) extracting an antigen according to claim 1 from prostatic tissue or separating said antigen from prostatic fluid; (b) separating said antigen from extraneous antigenic proteins to obtain a composition consisting essentially of said antigen; (c) immunizing animals with the resultant purified antigen to form antibodies specific thereto; and (d) recovering immunoprecipitating antibodies against said antigen from said animals.
Claim 10
Dependent Claim
19 words
A process according to claim 7, wherein said antigen is separated from extraneous antigenic protein material by salting out.
Claim 11
Dependent Claim
14 words
A process according to claim 7, wherein said antigen is obtained from seminal fluid.
Claim 12
Dependent Claim
65 words
A process preparing antibodies to antigens associated with prostatic tissue and fluid, which comprises: (a) cloning hybridoma cells capable of secreting said antibodies; (b) extracting antibodies according to claim 2 from said secretions; (c) separating said antibodies from extraneous antigenic proteins to obtain a composition consisting essentially of said antibodies; and (d) recovering an immunologically effective concentration and amount of said antibodies from said cells.
Claim 13
Dependent Claim
24 words
A process according to claim 10, wherein said hybridoma cells are prepared by fusing a nonsecretory myeloma cell line with primary mouse spleen cells.
Claim 14
Dependent Claim
75 words
A method for diagnosing carcinoma of the prostate, which comprises: (a) forming an immunoprecipitin complex between (i) an an antigen consisting essentially of the human prostate specific antigen of claim 1 associated with both normal and cancerous prostatic tissue which is distinct from prostatic acid phosphatase and (ii) immunoprecipitating antibodies thereto which are free of cross-reactivity against prostatic acid phosphatases and against antigens associated with other carcinomas; and (b) detecting the presence of said complex.
Claim 15
Dependent Claim
14 words
A method according to claim 12, wherein the complex is formed by countercurrent immunoelectrophoresis.
Claim 16
Dependent Claim
25 words
A method according to claim 12, wherein the complex is formed by immunologically reacting said antigen with antibodies thereto covalently bound to a water-insoluble carrier.
Claim 17
Dependent Claim
17 words
A method according to claim 12, wherein said complex is detected by radioimmunoassay or by enzyme-linked immunoassay.
Claim 18
Dependent Claim
53 words
A continuous murine cell line capable of producing monoclonal antibodies of the IgM isotype to the specific antigen of claim 1 under nutrient growth conditions, consisting essentially of a fused cell hybrid of: (a) primed antibody-producing cells with (b) myeloma cells capable of producing a homogenous population of immunoglobulin in the fused hybridoma.
Claim 19
Dependent Claim
16 words
A cell line according to claim 16, wherein the fused cell hybrid is ATCC HB 8051.
References Cited (Backward Citations)
US4331647A
Cited
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